Trifluoromethyl substituted perfluoro quinolizines

ABSTRACT

A perfluorobicyclo compound of the general formula: ##STR1## wherein either or both of Ring A and Ring B may optionally be substituted with lower perfluoroalkyl group(s) and m and n each represent 3 or 4.

This invention relates to novel perfluorobicyclo compounds useful asoxygen-carrying components for blood substitutes, oxygen-carryingtransfusions, and the like.

More specifically, this invention relates to perfluorobicyclo compoundsof the general formula: ##STR2## wherein either or both of Ring A andRing B may optionally be substituted with a lower perfluoroalkylgroup(s) and m and n each represent 3 or 4.

In the general formula (I), m and n each represent 3 or 4, and thus RingA and Ring B each represents a five-membered ring or a six-memberedring, and thus a condensed ring is formed from these two.

Examples of the condensed ring formed by Ring A and Ring B includeperfluor oderivatives of perhydroquinolizine, indolizine, pyrrolizine,etc.

Either or both of Ring A and Ring B may optionally be substituted withone or more, preferably b 1-3, lower perfluoroalkyl groups at anyposition. The perfluoroalkyl group as the substituent is either linearor branched, and examples of these groups include those having 1-4carbon atoms, such as perfluoromethyl group, perfluoroethyl group,perfluoro-n-propyl group, perfluoro-iso-propyl group, perfluoro-n-butylgroup, perfluoro-iso-butyl group, perfluoro-sec-butyl group andperfluoro-tert-butyl group, among which those having 1-3 carbon atomsare preferred. When two or more such substituents are present, they maybe different from each other.

The total carbon number in the compound of the general formula (I) isgenerally 8-12, preferably 9-11, more preferably 10.

The compound of the general formula (I) may be produced by fluorinatinga perhydrocompound corresponding to the compound (I). As thefluorination method, there may be mentioned, for example, knownfluorination methods such as direct fluorination method, cobaltfluorination method, electrolytic fluorination method, etc.

For the production of the compound (I) of this invention, theelectrolytic fluorination method is preferred, and this may be effectedby, for example, mixing and dissolving anhydrous hydrogen fluoride and aperhydro compound, as a starting material in an electrolytic cell, andthereafter effecting electrolysis. In said electrolysis, the voltage isgenerally 3-9 V, the anode current density is generally 1-300 A/dm², andthe bath temperature is generally 4°-10° C.

Since the thus formed compound of the general formula (I) is insolublein anhydrous hydrofluoric acid, it precipitates in the lower layer ofthe electrolytic cell.

Separation and purification of the compound (I) from said precipitatesmay be effected by, for example, adding a liquid mixture of an equalvolume of an aqueous alkali solution and an amine compound to therecovered precipitates, separating the lowest layer containing thecompound (I) (at that time, partially fluorinated compounds areseparated into the amine layer), washing it with an appropriate amountof aqueous acetone containing potassium iodide to remove compoundshaving fluorine atoms bound to nitrogen atoms, and further subjecting tofractional distillation to separate the compound (I).

Since, in addition to the capability of dissolving large quantities ofoxygen, the compound of the general formula (I) of this invention ismetabilically inert and also is rapidly eliminated from the body, forexample, an aqueous emulsion containing 5-50% (w/v), preferably 10-40%(w/v), of the compound (I) may be prepared and advantageously employedas oxygen-carrying components for blood substituents, oxygen-carryingtransfusions, etc. for warm-blooded animals including humans (e.g. dogs,cats, cows, horses, rats, mice, guinea pigs, etc.).

On preparing the aforesaid emulsion, an emulsifier such as polymericnonionic surfactants, phospholipids, etc., are employed in an amount of1-5% (w/v) each alone or in combination.

As the medium, a physiologically acceptable aqueous solution isemployed, and, if necesary, an isotonizing amount of an isotonizingagent such as glycerol may be added to isotonize the emulsion, and aplasma extender such as hydroxyethylstarch, dextran etc., may be addedto adjust the colloid osmotic pressure of the emulsion.

Thus, an aqueous emulsion may be prepared by homogenizing the variouscomponents as those described above to a particle size of 0.05-0.3μ,preferably to 0.2μ or less, using e.g. a high-pressure jet-typehomogenizer.

The perhydro compounds corresponding to the compound of the generalformula (I), as starting materials are substantially known compounds.

The present invention is further illustrated in more detail below withreference to Examples and Reference Examples, but the invention is notlimited thereto.

EXAMPLE 1

As an electrolytic cell, a tank (made of Monel metal (having a capacityof 1.5 l, containing electrodes made of nickel (purity of 99.6% orhigher) (6 anodes and 7 cathodes) alternately arranged with an electrodegap of 1.7-2.0 mm with an effective anode surface area of 10.5 dm² andfurther equipped with reflex condenser made of copper above the cell,was employed.

Into this electrolytic cell was introduced 1.2 l of anhydronshydrofluoric acid, and impurities (water and sulfuric acid) present invery small amounts were removed by the preliminary electrolysis.Thereafter, 0.85 mole (130 g) of 4-methyloctahydroquinolizine(4-methylperhydroquinolizine) was dissolved in the hydrofluoric acid,and while passing helium gas at a flow rate of 100 ml/min from the lowerpart of the cell, the electrolysis was effected with an anode currentdensity of 1.0-2.0 A/dm², a voltage of 4.0-6.2 V and a bath temperatureof 4°-10° C. The electrolysis was continued for 1051 A·hr until theelectrolytic voltage reached 9.0 V. Anhydrous hydrofluoric acid wasadditionally introduced 200 ml per 24 hours. The gas generated duringthe electrolysis was passed through an iron tube packed with sodiumfluoride pellets to remove the entrained anhydrous hydrofluoric acid, anthen led into a trap cooled with dry ice-acetone to liquefy and trap theproduct. On the other hand, the bath liquid in the electrolytic cellseparated into two layers, the upper layer containing hydrogen fluorideand the lower layer fluorobicyclo compounds. The lower layer wasseparated.

The liquid collected by cooling the above generated gas and the lowerlayer liquid of the electrolytic cell were combined, 70% KOH aqueoussolution and diisobutylamine were added in equal volumes, and refluxingwas conducted for 7 days. The perfluoro product was separated using aseparatory funnel, washed with 70% (w/v) acetone aqueous solutioncontaining 10% (w/v) of potassium iodide, then subjected to fractionaldistillation using an apparatus for fractional distillation equippedwith a spinning band column to obtainperfluoro-4-methyloctahydroquinolizine(perfluoro-4-methylquinolizine)(b.p. 145°-155° C./760 mm Hg). Said compound was confirmed to be thedesired compound perfluoro-4-methyloctahydroquinolidine also by theinfrared absorption spectrum, F nuclear magnetic resonance spectrum, andmass spectrum.

EXAMPLES 2-101

Perfluorobicyclo compounds corresponding to perhydro compounds used asstarting materials were obtained by the procedures similar to those inExample 1. These are sammarized in Table 1.

                                      TABLE 1    __________________________________________________________________________    Example                                  Boiling point    No.  Raw material compound                           Perfluorocompound (°C./760 mm    __________________________________________________________________________                                             Hg)    1    4-methylperhydroquinolizine                            ##STR3##         145-155     2   perhydroquinolizine                            ##STR4##         129-130     3   3-methylperhydroindolizine                            ##STR5##         125-135     4   2-methylperhydroindolizine                            ##STR6##         125-136     5   1-methylperhydroindolizine                            ##STR7##         125-135     6   8a-methylperhydroindolizine                            ##STR8##         125-136     7   8-methylperhydroindolizine                            ##STR9##         125-135     8   7-methylperhydroindolizine                            ##STR10##        125-135     9   5-methylperhydroindolizine                            ##STR11##        125-135     10  5-methylperhydroindolizine                            ##STR12##        125-136     11  3-methylperhydroquinolizine                            ##STR13##        145-155     12  2-methylperhydroquinolizine                            ##STR14##        145-155     13  1-methylperhydroquinolizine                            ##STR15##        145-155     14  9a-methylperhydroquinolizine                            ##STR16##        145-156     15  3-ethylperhydroindolizine                            ##STR17##        144-155     16  2-ethylperhydroindolizine                            ##STR18##        145-155     17  1-ethylperhydroindolizine                            ##STR19##        145-155     18  8a-ethylperhydroindolizine                            ##STR20##        145-156     19  8-ethylperhydroindolizine                            ##STR21##        145-155     20  7-ethylperhydroindolizine                            ##STR22##        145-155     21  6-ethylperhydroindolizine                            ##STR23##        145-155     22  5-ethylperhydroindolizine                            ##STR24##        145-155     23  2,3-dimethylperhydroindolizine                            ##STR25##        145-155     24  1,3-dimethylperhydroindolizine                            ##STR26##        145-156     25  3,8a-dimethylperhydroindolizine                            ##STR27##        145-155     26  3,8-dimethylperhydroindolizine                            ##STR28##        145-155     27  3,7-dimethylperhydroindolizine                            ##STR29##        145-155     28  3,6-dimethylperhydroindolizine                            ##STR30##        145-155     29  3,5-dimethylperhydroindolizine                            ##STR31##        145-156     30  1,2-dimethylperhydroindolizine                            ##STR32##        145-155     31  2,8a-dimethylperhydroindolizine                            ##STR33##        144-155     32  2,8-dimethylperhydroindolizine                            ##STR34##        145-156     33  2,7-dimethylperhydroindolizine                            ##STR35##        145-156     34  2,6-dimethylperhydroindolizine                            ##STR36##        144-155     35  2,5-dimethylperhydroindolizine                            ##STR37##        145-155     36  1,8a-dimethylperhydroindolizine                            ##STR38##        145-156     37  1,8-dimethylperhydroindolizine                            ##STR39##        145-156     38  1,7-dimethylperhydroindolizine                            ##STR40##        145-156     39  1,6-dimethylperhydroindolizine                            ##STR41##        144-155     40  1,5-dimethylperhydroindolizine                            ##STR42##        145-155     41  8,8a-dimethylperhydroindolizine                            ##STR43##        145- 155     42  7,8a-dimethylperhydroindolizine                            ##STR44##        145-155     43  6,8a-dimethylperhydroindolizine                            ##STR45##        145-156     44  5,8a-dimethylperhydroindolizine                            ##STR46##        145-156     45  7,8-dimethylperhydroindolizine                            ##STR47##        145-155     46  6,8-dimethylperhydroindolizine                            ##STR48##        145-155     47  5,9-dimethylperhydroindolizine                            ##STR49##        145-156     48  6,7-dimethylperhydroindolizine                            ##STR50##        145-155     49  5,6-dimethylperhydroindolizine                            ##STR51##        145-155     50  5,6-dimethylperhydroindolizine                            ##STR52##        145-155     51  3,3-dimethylperhydroindolizine                            ##STR53##        145-155     52  1,1-dimethylperhydroindolizine                            ##STR54##        145-155     53  8,8-dimethylperhydroindolizine                            ##STR55##        145-155     54  7,7-dimethylperhydroindolizine                            ##STR56##        145-155     55  6,6-dimethylperhydroindolizine                            ##STR57##        145-155     56  5,5-dimethylperhydroindolizine                            ##STR58##        145-155     57  3-propylperhydropyrrolizine                            ##STR59##        145-155     58  2-propylperhydropyrrolizine                            ##STR60##        145-156     59  7a-propylperhydropyrrolizine                            ##STR61##        145-156     60  1-propylperhydropyrrolizine                            ##STR62##        145-156     61  3-ethyl-2-methyperhydropyrrolizine                            ##STR63##        145-156     62  3-ethyl-1-methylperhydropyrrolizine                            ##STR64##        145-155     63  3-ethyl-7a-methylperhydropyrrolizine                            ##STR65##        145-156     64  3-ethyl-7-methylperhydropyrrolizine                            ##STR66##        145-156     65  3-ethyl-6-methylperhydropyrrolizine                            ##STR67##        145-155     66  3-ethyl-5-methylperhydropyrrolizine                            ##STR68##        145-156     67  2-ethyl-3-methylperhydropyrrolizine                            ##STR69##        145-155     68  2-ethyl-1-methylperhydropyrrolizine                            ##STR70##        145-156     69  2-ethyl-7a-methylperhydropyrrolizine                            ##STR71##        145-156     70  2-ethyl-7-methylperhydropyrrolizine                            ##STR72##        145-155     71  2-ethyl-6-methylperhydropyrrolizine                            ##STR73##        145-156     72  2-ethyl-5-methylperhydropyrrolizine                            ##STR74##        145-156     73  1-ethyl-3-methylperhydropyrrolizine                            ##STR75##        145-155     74  1-ethyl-2-methylperhydropyrrolizine                            ##STR76##        145-156     75  1-ethyl-7a-methylperhydropyrrolizine                            ##STR77##        145-157     76  1-ethyl-7-methylperhydropyrrolizine                            ##STR78##        145-156     77  1-ethyl-6-methylperhydropyrrolizine                            ##STR79##        145-155     78  1-ethyl-5-methylperhydropyrrolizine                            ##STR80##        145-156     79  7a-ethyl-3-methylperhydropyrrolizine                            ##STR81##        145-155     80  7a-ethyl-2-methylperhydropyrrolizine                            ##STR82##        145-156     81  7a-ethyl-1-methylperhydropyrrolizine                            ##STR83##        145-156     82  1,2,3-trimethylperhydropyrrolizine                            ##STR84##        145-155     83  2,3,7a-trimethylperhydropyrrolizine                            ##STR85##        145-155     84  2,3,7-trimethylperhydropyrrolizine                            ##STR86##        145-155      85? 2,3,6-trimethylperhydropyrrolizine?     ##STR87##

REFERENCE EXAMPLE 1

400 g of Vitelline phospholipid was added to 8.5 l of lactic acid addedRinger's solution, and stirred by a mixer to prepare a coarse emulsion,then 2.5 kg of perfluoro-4-methyloctahydroquinolizine was added theretoand stirred vigorously again by the mixer to prepare a coarse emulsion.This coarse emulsion was placed in a liquid tank of a jet emulsifier(manufactured by Manton-Gaulin Co.) and circulated while maintaining theliquid temperature at 50°±5° C. to effect emulsification. Theconcentration of perfluoro-4-methyloctahydroquinolidine in the obtainedemulsion was 27.3% (w/v). The particle diameter as measured by thecentrifugal sedimentation method was 0.05-0.25μ. This emulsion wasallotted into vials for injection, stoppered and thermally sterilized ina rotary sterilizer, but there was no significant increase in theparticle diameter observed.

REFERENCE EXAMPLE 2

An emulsion was obtained by procedures similar to those in ReferenceExample 1 except that perfluoro-4-methyloctahydroquinolidine wasreplaced by perpluoro-4-ethyloctahydroquinolidine. The particle diameterof the thus obtained emulsion was 0.0.5-0.25μ.

EXPERIMENTAL EXAMPLE 1 Stability of Emulsions

Water was added to 20 g of each perfluorobicyclo compound selected inthe present invention and 4 g of vitelline phospholipid to make thetotal volume 200 ml, and emulsification was effected using aManton-Gaulin emulsifier as used above under nitrogen stream at 200-600kg/cm² while maintaining the liquid temperature at 40°-45° C. Eachobtained emulsion was filtered through a 0.65μ membrane filter, allottedinto 20 ml-capacity vials, and, after replacing the atmosphere bynitrogen gas, thermally treated at 100° C. for 30 minutes, followed bystoring at 4° C. of room temperature to examine the stability. Theparticle diameter of the emulsion was measured by the centrifugalsedimentation method by Yokoyama et al. [Chem. Pharm. Bull. 22 (12) 2966(1974)], and from the obtained data, the average particle diameterdistribution was calculated using a microcomputer.

Thus, the particle diameter distributions of each perfluoro-carbonemulsion before and after heating, and after heating and storing at 4°C. and room temperature (15°-28° C.) are shown in Tables 2 and 3. As isevident from the results, the emulsions according to the presentinvention are very stable against heating and the influence on theaverage particle diameter due to heating was not observed at all.Further, when stored at 4° C. after heating, there was no increase inthe average particle diameter observed even after 5 months.

                                      TABLE 2    __________________________________________________________________________    Stability of Perfluoro-4-methyloctahydroquinolizine Emulsion                 Average                 Particle                       Distribution of Particle Diameter                 Diameter,                       (wt %)                 μ  <0.1μ                           0.1-0.2μ                                0.2-0.3μ                                     >0.3μ    __________________________________________________________________________    Before heating                 0.122 39.0                           49.9 14.2 0.9    Immediately after heating                 0.122 35.2                           56.3 8.5  0    After 2 weeks    at 4° C.                 0.116 37.1                           59.4 3.5  0    at R.T.*     0.127 31.2                           60.1 8.7  0    After 4 weeks    at 4° C.                 0.122 33.0                           61.5 5.5  0    at R.T.*     0.114 31.3                           68.6 0.2  0    After 5 months    at 4° C.                 0.135 25.5                           64.2 10.4 0    __________________________________________________________________________     *R.T. = Room Temperature

                                      TABLE 3    __________________________________________________________________________    Stability of Perfluoro-4-ethyloctahydroquinolizine Emulsion                 Average                 Particle                       Distribution of Particle Diameter                 Diameter,                       (wt %)                 μ  <0.1μ                           0.1-0.2μ                                0.2-0.3μ                                     >0.3μ    __________________________________________________________________________    Before heating                 0.122 38.0                           51.2 10.8 0    Immediately after heating                 0.122 34.3                           57.5 8.2  0    After 2 weeks    at 4° C.                 0.118 37.1                           61.0 1.9  0    at R.T.*     0.128 31.3                           63.5 5.2  0    After 4 weeks    at 4° C.                 0.123 38.5                           60.2 1.3  0    at R.T.*     0.119 34.1                           60.5 5.4  0    After 5 months    at 4° C.                 0.126 37.9                           58.1 4.0  0    __________________________________________________________________________     *R.T. = Room Temperature

EXPERIMENTAL EXAMPLE 2 Acute Toxicity Test

The acute toxicity test on the preparations of the present invention wascarried out using the preparations of the present invention shown inTable 4 which had been physiologically isotonized. The test animals usedwere Wister-strain male rats (weighing 100-120 g). The emulsion wasintravenously administered and the animals were observed for one weekafter the administration.

The results are such that with either emulsion containingperfluoro-4-methyloctahydroquinolizine orperfluoro-4-ethyloctahydroquinolizine, there was no death case at 100ml/kg-body weight and thus their toxicity are very small.

                  TABLE 4    ______________________________________                              Ratio    Composition               % (w/v)    ______________________________________    Oil      Perfluorobicyclo Compound                                  30    Component             Emulsify- Vitelline Phospholipid                                      4.0    (9 vol)  ing Agent    Electro-           NaCl           6.00    lyte               NaHCO.sub.3    2.1    (1 vol)            KCl            0.336                       MgCl.sub.2.6H.sub.2 O                                      0.427                       CaCl.sub.2.2H.sub.2 O                                      0.356                       D-Glucose      1.802           pH      8.0    ______________________________________

EXPERIMENTAL EXAMPLE 3 Distribution of Perfluoro-compound in Organs

Using Wister-strain male rats weighing 120-130 g, the emulsion preparedin Reference Example 1 was administered into the tail vein [at 4 g/kg asperfluoro-4-methyloctahydroquinolidine], and for a period of 3 monthsafter the administration, the content of said compound in the liver,spleen and fat tissues due to uptake were measured by means of gaschromatography.

The content of perfluoro-4-methyloctahydroperhydroquinolizine uptake ineach organ 1, 2 and 4 weeks and 3 months after the administration areshown in Table 5. The compound was taken up in greater amounts by thereticulo-endothelial organs shortly after the administration, but soondisappeared rapidly. There was no evidence of adverse influence on theliver or spleen organ.

As a result, the half-life of perfluoro-4-methylperhydroquinolizine wascalculated to be 7.33 days.

                  TABLE 5    ______________________________________                           Residual Rate of               Time after the                           perfluorobicyclo    Organ      administration                           compound, %    ______________________________________    Liver      1 Week      19.92               2 Weeks     8.66               4 Weeks     1.88               3 Months    0.30    Spleen     1 Week      11.61               2 Weeks     9.33               4 Weeks     2.45               3 Months    0.09    ______________________________________

EXPERIMENTAL EXAMPLE 4 Anatominal Remarks

Wister-strain male rats weighing 120-130 g were administered with 4 g/kgof the perfluorobicyclo-compound emulsion prepared in Reference Example1 or 2, and the dissected organs were observed for a period of 3 monthsafter the administration, and further the organs (liver and spleen) wereweighed, to determine the weight relatige to the body weight.

One, 2 and 4 weeks and 3 months after the administration of theemulsion, the important organs, i.e. the lung, liver and spleen wereobserved, to find no evidence of the influence on the organs by eithersaid compound because of their rapid elimination.

What is claimed is:
 1. A perfluorobicyclo compound of the generalformula: ##STR88## wherein either or both of Ring A and Ring B mayoptionally be substituted with perfluoroalkyl group(s) having from 1 to3 carbom atoms, and the total number of carbon atoms in the compound ofthe above formula is 10 to
 12. 2. A compound according to claim 1 whichis perfluoro-4-methyloctahydroquinolizine.
 3. A compoundd according toclaim 1 which is perfluoro-2-methyloctahydroquinolizine.
 4. A compoundaccording to claim 1 which is perfluoro-1-methyloctahydroquinolizine. 5.A compound according to claim 1 which isperfluoro-9a-methyloctahydroquinolizine.
 6. A compound according toclaim 1 which is perfluoro-4-ethyloctahydroquinolizine.
 7. A compoundaccording to claim 1 having 10 to 11 carbon atoms.
 8. A compoundaccording to claim 1 substantially free from other isomers.